Keratin as an effective coating material for in vitro stem cell culture, induced differentiation and wound healing assays

Hina F Bhat; Nuzhat Amin; Zarka Nasir; Saba Nazir; Zuhaib F Bhat; Abrar A Malik; Nazir A Ganai; S Mudasir Andrabi; Riaz A Shah; Rana Muhammad Aadil; Asif H Sofi; Gholamreza Abdi

doi:10.1016/j.heliyon.2024.e27197

Keratin as an effective coating material for in vitro stem cell culture, induced differentiation and wound healing assays

Heliyon. 2024 Feb 29;10(5):e27197. doi: 10.1016/j.heliyon.2024.e27197. eCollection 2024 Mar 15.

Authors

Affiliations

¹ Division of Animal Biotechnology, SKUAST-Kashmir, Srinagar, J&K, 190006, India.
² Department of Biotechnology, BGSB University, Rajouri, J&K, India.
³ Division of Livestock Products Technology, SKUAST-Jammu, J&K, India.
⁴ National Institute of Food Science and Technology, University of Agriculture, Faisalabad, Pakistan.
⁵ Division of Livestock Products Technology, SKUAST-Kashmir, Srinagar, J&K, 190006, India.
⁶ Department of Biotechnology, Persian Gulf Research Institute, Persian Gulf University, 75169, Iran.

Abstract

The utilization of stem cells in tissue engineering holds great promise as efficient tools for tissue regeneration and in treating numerous musculoskeletal diseases. However, several limiting factors, such as precise delivery and control of differentiation of these stem cells as well as mimicking the microenvironment required to modulate stem cell behaviour in-vivo, have given rise to an urgent need for the development of new biomaterials which could be tailored to enhance cell renewal and/or direct cell fates. Keratin-rich biological materials offer several advantages, such as biocompatibility, tailorable mechanical properties, huge bioavailability, non-toxicity, non-immunogenic, and intrinsic tissue repair and/or regeneration capabilities, which makes them highly valued. In the present work, we report the preparation of keratin-based bio-materials from goat hair waste and its effectiveness as a coating material for in vitro culture and induced differentiation of mesenchymal stem cells (MSC's) and primary goat fibroblast cells. Since no known keratinase enzymes are expressed as such in human and/or animal systems, these keratin biomaterials could be used to slow the rate of degradation and deliver keratin-loaded stem cell scaffolds to induce their directed differentiation in vivo. The generated keratin materials have been characterized for surface morphology, protein structures, size and other properties using SDS-PAGE, LC/MS-MS, SEM, FTIR etc. Also, in vitro cell culture assays such as cell adhesion, viability using MTT, live dead assays, differentiation assays and in vitro scratch/wound healing assays were performed. Our results provide important data supporting tissue engineering applications of these keratinous biomaterials by combining the unique biological characteristics of goat hair-derived keratin material with the regenerative power of stem cells and their combinatorial use in applications such as disease treatment and injury repair as well as their use in the preparation of wound healing products, such as dressings and bandages, for management of clinical care in animals.

Keywords: Cell culture assays; FTIR; Keratin; Mesenchymal stem cells (MSC's); SDS-PAGE; SEM.